Semisynthetic fluorescent pH sensors for imaging exocytosis and endocytosis

Nat Commun. 2017 Nov 10;8(1):1412. doi: 10.1038/s41467-017-01752-5.

Abstract

The GFP-based superecliptic pHluorin (SEP) enables detection of exocytosis and endocytosis, but its performance has not been duplicated in red fluorescent protein scaffolds. Here we describe "semisynthetic" pH-sensitive protein conjugates with organic fluorophores, carbofluorescein, and Virginia Orange that match the properties of SEP. Conjugation to genetically encoded self-labeling tags or antibodies allows visualization of both exocytosis and endocytosis, constituting new bright sensors for these key steps of synaptic transmission.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques / methods
  • Drug Design
  • Endocytosis*
  • Exocytosis*
  • Fluoresceins / chemical synthesis
  • Fluoresceins / chemistry
  • Fluorescent Dyes* / chemical synthesis
  • Fluorescent Dyes* / chemistry
  • Green Fluorescent Proteins / chemistry
  • Hippocampus / cytology
  • Hippocampus / metabolism
  • Hydrogen-Ion Concentration
  • Luminescent Proteins / chemistry
  • Neurons / metabolism
  • PC12 Cells
  • Rats
  • Red Fluorescent Protein
  • Synaptic Transmission
  • Synaptic Vesicles / physiology

Substances

  • Fluoresceins
  • Fluorescent Dyes
  • Luminescent Proteins
  • PHluorin
  • Green Fluorescent Proteins